4-Bromo or 4-iodo phenylamino benzhydroxamic acid derivatives and their use as MEK inhibitors

ABSTRACT

Phenylamino benzhydroxamic acid derivatives of formula (I) where R 1 , R 2 , R 3 , R 4 , R 5 , and R 6  are hydrogen or substituent groups such as alkyl, and where R 7  is hydrogen or an organic radical, are potent inhibitors of MEK and, as such, are effective in treating cancer and other proliferative diseases such as psoriasis and restenosis.

This application is a continuation application of U.S. Ser. No.09/462,239 filed Jan. 4, 2000, now abandoned which is a 371 applicationof PCT/US98/13106 filed Jun. 24, 1998, which claims the benefit ofpriority to U.S. provisional application Serial No. 60/051,440 filedJul. 1, 1997.

FIELD OF THE INVENTION

This invention provides certain hydroxamic acid derivatives ofanthranilic acids which inhibit certain dual specificity kinase enzymesinvolved in proliferative diseases such as cancer and restenosis.

BACKGROUND OF THE INVENTION

Proliferative diseases are caused by a defect in the intracellularsignaling system, or the signal transduction mechanism of certainproteins. Cancer, for example, is commonly caused by a series of defectsin these signaling proteins, resulting from a change either in theirintrinsic activity or in their cellular concentrations. The cell mayproduce a growth factor that binds to its own receptors, resulting in anautocrine loop, which continually stimulates proliferation. Mutations oroverexpression of intracellular signaling proteins can lead to spuriousmitogenic signals within the cell. Some of the most common mutationsoccur in genes encoding the protein known as Ras, which is a G-proteinthat is activated when bound to GTP, and inactivated when bound to GDP.

The above mentioned growth factor receptors, and many other mitogenicreceptors, when activated, lead to Ras being converted from theGDP-bound state to the GTP-bound state. This signal is an absoluteprerequisite for proliferation in most cell types. Defects in thissignaling system, especially in the deactivation of the Ras.GTP complex,are common in cancers, and lead to the signaling cascade below Ras beingchronically activated.

Activated Ras leads in turn to the activation of a cascade ofserine/threonine kinases. One of the groups of kinases known to requirean active Ras.GTP for its own activation is the Raf family. These inturn activate MEK, which then activates MAP kinase. Activation of MAPkinase by mitogens appears to be essential for proliferation, andconstitutive activation of this kinase is sufficient to induce cellulartransformation. Blockade of downstream Ras signaling, for example by useof a dominant negative Raf-1 protein, can completely inhibitmitogenesis, whether induced from cell surface receptors or fromoncogenic Ras mutants. Although Ras is not itself a protein kinase, itparticipates in the activation of Raf and other kinases, most likelythrough a phosphorylation mechanism. Once activated, Raf and otherkinases phosphorylate MEK on two closely adjacent serine residues, S²¹⁸and S²²² in the case of MEK-1, which are the prerequisite for activationof MEK as a kinase. MEK in turn phosphorylates MAP kinase on both atyrosine, Y¹⁸⁵, and a threonine residue, T¹⁸³, separated by a singleamino acid. This double phosphorylation activates MAP kinase at least100-fold, and it can now catalyze the phosphorylation of a large numberof proteins, including several transcription factors and other kinases.Many of these MAP kinase phosphorylations are mitogenically activatingfor the target protein, whether it be another kinase, a transcriptionfactor, or other cellular protein. MEK is also activated by severalkinases other than Raf-1, including MEKK, and itself appears to be asignal integrating kinase. As far as is currently known, MEK is highlyspecific for the phosphorylation of MAP kinase. In fact, no substratefor MEK other than MAP kinase has been demonstrated to date, and MEKdoes not phosphorylate peptides based on the MAP kinase phosphorylationsequence, or even phosphorylate denatured MAP kinase. MEK also appearsto associate strongly with MAP kinase prior to phosphorylating it,suggesting that phosphorylation of MAP kinase by MEK may require a priorstrong interaction between the two proteins. Both this requirement andthe unusual specificity of MEK are suggestive that it may have enoughdifference in its mechanism of action to other protein kinases thatselective inhibitors of MEK, possibly operating through allostericmechanisms rather than through the usual blockade of the ATP bindingsite, may be found.

This invention provides compounds which are highly specific inhibitorsof the kinase activity of MEK. Both in enzyme assays and whole cells,the compounds inhibit the phosphorylation of MAP kinase by MEK, thuspreventing the activation of MAP kinase in cells in which the Rascascade has been activated. The results of this enzyme inhibitioninclude a reversal of transformed phenotype of some cell types, asmeasured both by the ability of the transformed cells to grow in ananchorage-independent manner and by the ability of some transformed celllines to proliferate independently of external mitogens.

The compounds provided by this invention are phenylamino benzhydroxamicacid derivatives in which the phenyl ring is substituted at the4-position with bromo or iodo. U.S. Pat. No. 5,155,110 discloses a widevariety of fenamic acid derivatives, including certain phenylaminobenzhydroxamic acid derivatives, as anti-inflammatory agents. Thereference fails to describe the compound of this invention or theirkinase inhibitory activity.

SUMMARY OF THE INVENTION

This invention provides 4-bromo and 4-iodo phenylamino benzhydroxamicacid derivatives which are kinase inhibitors and as such are useful fortreating proliferative diseases such as cancer, psoriasis, andrestenosis. The compounds are defined by Formula I

wherein:

R₁ is hydrogen, hydroxy, C₁-C₈ alkyl, C₁-C₈ alkoxy, halo,trifluoromethyl, or CN;

R₂ is hydrogen;

R₃, R⁴ and R₅ independently are hydrogen, hydroxy, halo,trifluoromethyl, C₁-C₈ alkyl, C₁-C₈ alkoxy, nitro, CN, or (O orNH)_(m)—(CH₂)_(n)—R₉, where R₉ is hydrogen, hydroxy, CO₂H or NR₁₀R₁₁;

n is 0 to 4;

m is 0 or 1;

R₁₀ and R₁₁ independently are hydrogen or C₁-C₈ alkyl, or taken togetherwith the nitrogen to which they are attached can complete a 3- to10-member cyclic ring optionally containing one, two, or threeadditional heteroatoms selected from O, S, NH, or N—C₁-C₈ alkyl;

R₆ is hydrogen, C₁-C₈ alkyl,

alkyl, aryl, aralkyl, or C₃-C₁₀ cycloalkyl;

R₇ is hydrogen, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₁₀(cycloalkyl or cycloalkyl optionally containing a heteroatom selectedfrom O, S, or NR₉); or R₆ and R₇ taken together with the N—O to whichthey are attached can complete a 5- to 10-membered cyclic ring,optionally containing one, two, or three additional heteroatoms selectedfrom O, S, or NR₁₀R₁₁;

and wherein any of the foregoing alkyl, alkenyl, and alkynyl groups canbe unsubstituted or substituted by cycloalkyl (or cycloalkyl optionallycontaining a heteroatom selected from O, S, or NR₉), aryl, aryloxy,heteroaryl, or heteroaryloxy.

Preferred compounds have Formula II

where R₁, R₃, R₄, R₅, R₆, and R₇ are as defined above. Especiallypreferred are compounds wherein R₁ is methyl or halo, and R₃, R₄, and R₅are halo such as fluoro or bromo.

Another preferred group of compounds have Formula III

wherein R₁, R₃, R₄, R₅, and R₇ are as defined above.

The most preferred compounds are those wherein R₁ is methyl or halo suchas F, Br, Cl, and I, R₃ is hydrogen or halo such as fluoro, R₄ is halosuch as fluoro, and R₅ is hydrogen or halo such as fluoro or bromo. Suchcompounds have the formulas

Specific compounds provided by the invention include the following:

3,4,5-Trifluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide;

5-Chloro-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide;

5-Bromo-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide;

N-Hydroxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzamide;

3,4,5-Trifluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide;

5-Chloro-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide;

5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide;

2-(2-Fluoro-4-iodo-phenylamino)-N-hydroxy-4-nitro-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-3,4,5-trifluoro-N-hydroxy-benzamide;

5-Chloro-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide;

5-Bromo-2-(2-bromo-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-N-hydroxy-4-methyl-benzamide;

2-(2-Bromo-4-iodo-phenylamino)-3,4,5-trifluoro-N-hydroxy-benzamide;

2-(2-Bromo-4-iodo-phenylamino)-5-chloro-3,4-difluoro-N-hydroxy-benzamide;

2-(2-Bromo-4-iodo-phenylamino)-N-hydroxy-4-nitro-benzamide;

4-Fluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide;

3,4-Difluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N-hydroxy-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide;

2-(2-Bromo-4-iodo-phenylamino)-4-fluoro-N-hydroxy-benzamide;

2-(2-Bromo-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide;

N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide;

5-Chloro-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide;

5-Bromo-N-cyclopropylmethoxy-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-benzamide;

N-Cyclopropylmethoxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzamide;

N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(2-fluoro-4-iodo-phenylamino)-benzamide;

5-Chloro-N-cyclopropylmethoxy-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-benzamide;

5-Bromo-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide;

N-Cyclopropylmethoxy-2-(2-fluoro-4-iodo-phenylamino)-4-nitro-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-benzamide;

5-Chloro-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide;

5-Bromo-2-(2-bromo-4-iodo-phenylamino)-N-ethoxy-3,4-difluoro-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-N-ethoxy-4-nitro-benzamide;

2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-benzamide;

2-(2-Bromo-4-iodo-phenylamino)-5-chloro-N-cyclopropylmethoxy-3,4-difluoro-benzamide

2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-nitro-benzamide;

N-Cyclopropylmethoxy-4-fluoro-2-(2-fluoro-4-iodo-phenylamino)-benzamide;

N-Cyclopropylmethoxy-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-fluoro-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide;

2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-4-fluoro-benzamide;

2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide;

4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-N-isopropyl-benzamide;

N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide;

4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-N-methyl-benzamide;

4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-N-hydroxy-4-nitro-benzamide;

3,4-Difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide;

2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N-hydroxy-benzamide (HCl salt);

2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N-(tetrahydro-pyran-2-yloxy)-benzamide;

3,4-Difluoro-2-(2-chloro-4-iodo-phenylamino)-N-cyclobutylmethoxy-benzamide;

5-Bromo-2-(2-chloro-4-iodo-phenylamino)-N-(2-dimethylamino-ethoxy)-3,4-difluoro-benzamidemonohydrochloride salt;

5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide;

3,4-Difluoro-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-benzamide;

5-Bromo-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide;

5-Bromo-N-cyclohexylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide;

5-Bromo-N-cyclopentylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide;and

5-Bromo-N-cyclobutylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide.

This invention also provides pharmaceutical formulations comprising acompound of Formula I together with a pharmaceutically acceptableexcipient, diluent, or carrier. Preferred formulations include any ofthe foregoing preferred compounds together with an excipient, diluent,or carrier.

The compounds of Formula I are potent and selective inhibitors of kinaseenzymes, particularly MEK₁ and MEK₂. They are, therefore, useful totreat subjects suffering from cancer and other proliferative diseasessuch as psoriasis, restenosis, autoimmune disease, and atherosclerosis.The compounds are especially well-suited to treat cancers such as breastcancer, colon cancer, prostate cancer, skin cancer, and pancreaticcancer. The compounds can also be used to treat stroke, diabetes,hepatomegaly, cardiomegaly, Alzheimer's disease, cystic fibrosis, andviral disease. The invention provides a method of inhibiting MEK enzymesand the foregoing diseases by administering to a subject an effectiveamount of a compound of Formula I.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the term “aryl” means a cyclic, bicyclic, or tricyclicaromatic ring moiety having from five to twelve carbon atoms. Examplesof typical aryl groups include phenyl, naphthyl, and fluorenyl. The arylmay be substituted by one, two, or three groups selected from fluoro,chloro, bromo, iodo, alkyl, hydroxy, alkoxy, nitro, or amino. Typicalsubstituted aryl groups include 3-fluorophenyl, 3,5-dimethoxyphenyl,4-nitronaphthyl, 2-methyl-4-chloro-7-aminofluorenyl, and the like.

The term “aryloxy” means an aryl group bonded through an oxygen atom,for example phenoxy, 3-bromophenoxy, naphthyloxy, and4-methyl-1-fluorenyloxy.

“Heteroaryl” means a cyclic, bicyclic, or tricyclic aromatic ring moietyhaving from four to eleven carbon atoms and one, two, or threeheteroatoms selected from O, S, or N. Examples include furyl, thienyl,pyrrolyl, pyrazolyl, triazolyl, thiazolyl, xanthenyl, pyronyl, indolyl,pyrimidyl, naphthyridyl, pyridyl, and triazinyl. The heteroaryl groupscan be unsubstituted or substituted by one, two, or three groupsselected from fluoro, chloro, bromo, iodo, alkyl, hydroxy, alkoxy,nitro, or amino. Examples of substituted heteroaryl groups includechloropyranyl, methylthienyl, fluoropyridyl, amino-1,4-benzisoxazinyl,nitroisoquinolinyl, and hydroxyindolyl.

The heteroaryl groups can be bonded through oxygen to make heteroaryloxygroups, for example thienyloxy, isothiazolyloxy, benzofuranyloxy,pyridyloxy, and 4-methylisoquinolinyloxy.

The term “C₁-C₈ alkyl” means straight and branched chain aliphaticgroups having from one to eight carbon atoms. Typical C₁-C₈ alkyl groupsinclude methyl, ethyl, isopropyl, tert.-butyl, 2,3-dimethylhexyl, and1,1-dimethylpentyl. The alkyl groups can be unsubstituted or substitutedby cycloalkyl, cycloalkyl containing a heteroatom selected from O, S, orNR₉, aryl, aryloxy, heteroaryl, or heteroaryloxy, as those terms aredefined above. Examples of aryl and aryloxy substituted alkyl groupsinclude phenylmethyl, 2-phenylethyl, 3-chlorophenylmethyl,1,1-dimethyl-3-(2-nitrophenoxy)butyl, and 3,4,5-trifluoronaphthylmethyl.Examples of alkyl groups substituted by a heteroaryl or heteroaryloxygroup include thienylmethyl, 2-furylethyl, 6-furyloxyoctyl,4-methylquinolyloxymethyl, and 6-isothiazolylhexyl. Cycloalkylsubstituted alkyl groups include cyclopropylmethyl, 2-cyclopentylethyl,2-piperidin-1-ylethyl, 3-(tetrahydropyran-2-yl)propyl, andcyclobutylmethyl.

“C₂-C₈ Alkenyl” means a straight or branched carbon chain having one ormore double bonds. Examples include but-2-enyl, 2-methyl-prop-2-enyl,1,1-dimethyl-hex-4-enyl, 3-ethyl-4-methyl-pent-2-enyl, and3-isopropyl-pent-4-enyl. The alkenyl groups can be substituted witharyl, aryloxy, heteroaryl, or heteroyloxy, for example3-phenylprop-2-enyl, 6-thienyl-hex-2-enyl, 2-furyloxy-but-2-enyl, and4-naphthyloxy-hex-2-enyl.

“C₂-C₈ Alkynyl” means a straight or branched carbon chain having fromtwo to eight carbon atoms and at least one triple bond. Typical alkynylgroups include prop-2-ynyl, 2-methyl-hex-5-ynyl,3,4-dimethyl-hex-5-ynyl, and 2-ethyl-but-3-ynyl. The alkynyl groups canbe substituted by aryl, aryloxy, heteroaryl, or heteroaryloxy, forexample 4-(2-fluorophenyl)-but-3-ynyl, 3-methyl-5-thienylpent-4-ynyl,3-phenoxy-hex-4-ynyl, and 2-furyloxy-3-methyl-hex-4-ynyl.

The alkenyl and alkynyl groups can have one or more double bonds ortriple bonds, respectively, or a combination of double and triple bonds.For example, typical groups having both double and triple bonds includehex-2-en-4-ynyl, 3-methyl-5-phenylpent-2-en-4-ynyl, and3-thienyloxy-hex-3-en-5-ynyl.

The term “C₃-C₁₀ cycloalkyl” means a non-aromatic ring or fused ringscontaining from three to ten carbon atoms. Examples include cyclopropyl,cyclobutyl, cyclopenyl, cyclooctyl, bicycloheptyl, adamantyl, andcyclohexyl. The ring can optionally contain a heteroatom selected fromO, S, or NR₉. Such groups include tetrahydrofuryl, tetrahydropyrrolyl,octahydrobenzofuranyl, octahydroindolyl, and octahydrobenzothiofuranyl.

R₃, R₄, and R₅ can include groups defined by the term (O orNH)_(m)—(CH₂)_(n)—R₉. Examples of such groups are aminomethyl,2-aminoethyl, 2-aminoethylamino, 3-aminopropoxy, N,N-diethylamino,3-(N-methyl-N-isopropylamino)-propylamino, 2-(N-acetylamino)-ethoxy,4-(N-dimethylaminocarbonylamino)-butoxy, and3-(N-cyclopropylamino)-propoxy.

The 4-bromo and 4-iodo phenylamino benzhydroxamic acid derivatives ofFormula I can be prepared from commercially available starting materialsutilizing synthetic methodologies well-known to those skilled in organicchemistry. A typical synthesis is carried out by reacting a 4-bromo or4-iodo aniline with a benzoic acid having a leaving group at the2-position to give a phenylamino benzoic acid, and then reacting thebenzoic acid phenylamino derivative with a hydroxylamine derivative.This process is depicted in Scheme 1.

where L is a leaving group, for example halo such as fluoro, chloro,bromo or iodo, or an activated hydroxy group such as a diethylphosphate,trimethylsilyloxy, p-nitrophenoxy, or phenylsulfonoxy.

The reaction of the aniline derivative and the benzoic acid derivativegenerally is accomplished by mixing the benzoic acid with an equimolarquantity or excess of the aniline in an unreactive organic solvent suchas tetrahydrofuran, or toluene, in the presence of a base such aslithium diisopropylamide, n-butyl lithium, sodium hydride, and sodiumamide. The reaction generally is carried out at a temperature of about−78° C. to about 25° C., and normally is complete within about 2 hoursto about 4 days. The product can be isolated by removing the solvent,for example by evaporation under reduced pressure, and further purified,if desired, by standard methods such as chromatography, crystallization,or distillation.

The phenylamino benzoic acid next is reacted with a hydroxylaminederivative HNR₆OR₇ in the presence of a peptide coupling reagent.Hydroxylamine derivatives that can be employed include methoxylamine,N-ethyl-isopropoxy amine, and tetrahydro-oxazine. Typical couplingreagents include 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ),1,3-dicyclohexylcarbodiimide (DCC), bromo-tris(pyrrolidino)-phosphoniumhexafluorophosphate (PyBrOP) and (benzotriazolyloxy)tripyrrolidinophosphonium hexafluorophosphate (PyBOP). The phenylamino benzoic acidand hydroxylamino derivative normally are mixed in approximatelyequimolar quantities in an unreactive organic solvent such asdichloromethane, tetrahydrofuran, chloroform, or xylene, and anequimolar quantity of the coupling reagent is added. A base such astriethylamine or diisopropylethylamine can be added to act as an acidscavenger if desired. The coupling reaction generally is complete afterabout 10 minutes to 2 hours, and the product is readily isolated byremoving the reaction solvent, for instance by evaporation under reducedpressure, and purifying the product by standard methods such aschromatography or crystallizations from solvents such as acetone,diethyl ether, or ethanol.

An alternative method for making the invention compounds involves firstconverting a benzoic acid to a hydroxamic acid derivative, and thenreacting the hydroxamic acid derivative with an aniline. This syntheticsequence is depicted in Scheme 2.

where L is a leaving group. The general reaction conditions for both ofthe steps in Scheme 2 are the same as those described above for Scheme1.

Yet another method for making invention compounds comprises reacting aphenylamino benzhydroxamic acid with an ester forming group as depictedin Scheme 3.

where L is a leaving group such as halo, and a base is triethylamine ordiisopropylamine.

The synthesis of invention compounds of Formula I is further illustratedby the following detailed examples.

EXAMPLE 1

4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

(a) Preparation of 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

To a stirred solution containing 3.16 g (0.0133 mol) of2-amino-5-iodotoluene in 5 mL of tetrahydrofuran at −78° C. was added 10mL (0.020 mol) of a 2.0 M lithium diisopropylamide intetrahydrofuran/heptane/ethylbenzene (Aldrich) solution. The resultinggreen suspension was stirred vigorously for 15 minutes, after which timea solution of 1.00 g (0.00632 mol) of 2,4-difluorobenzoic acid in 10 mLof tetrahydrofuran was added. The reaction 2,4-difluorobenzoic acid in10 mL of tetrahydrofuran was added. The reaction temperature was allowedto increase slowly to room temperature, at which temperature the mixturewas stirred for 2 days. The reaction mixture was concentrated byevaporation of the solvent under reduced pressure. Aqueous HCl (10%) wasadded to the concentrate, and the solution was extracted withdichloromethane. The organic phase was dried (MgSO₄) and thenconcentrated over a steambath to low volume (10 mL) and cooled to roomtemperature. The off-white fibers which formed were collected by vacuumfiltration, rinsed with hexane, and dried in a vacuum-oven (76° C.; ca.10 mm of Hg) to afford 1.10 g (47%) of the desired material; mp224-229.5° C.;

¹H NMR (400 MHz, DMSO): δ 9.72 (s, 1H), 7.97 (dd, 1H, J=7.0, 8.7 Hz),7.70 (d, 1H, J=1.5 Hz), 7.57 (dd, 1H, J=8.4, 1.9 Hz), 7.17 (d, 1H, J=8.2Hz), 6.61-6.53 (m, 2H), 2.18 (s, 3H);

¹³C NMR (100 MHz, DMSO): δ 169.87, 166.36 (d, J_(C-F)=249.4 Hz), 150.11(d, J_(C-F)=11.4 Hz), 139.83, 138.49, 136.07, 135.26 (d, J_(C-F)=11.5Hz), 135.07, 125.60, 109.32, 104.98 (d, J_(C-F)=21.1 Hz), 99.54 (d,J_(C-F)=26.0 Hz), 89.43, 17.52;

¹⁹F NMR (376 MHz, DMSO): δ−104.00 to −104.07 (m);

IR (KBr) 1670 (C═O stretch)cm⁻¹;

MS (CI) M+1=372.

Analysis calculated for C₁₄H₁₁FINO₂:

C, 45.31; H, 2.99; N, 3.77.

Found: C, 45.21; H, 2.77; N, 3.64.

(b) Preparation of4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

To a stirred solution of4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid (0.6495 g,0.001750 mol), O-(tetrahydro-2H-pyran-2-yl)-hydroxylamine (0.2590 g,0.002211 mol), and diisopropylethylamine (0.40 mL, 0.0023 mol) in 31 mLof an equivolume tetrahydrofuran-dichloromethane solution was added 1.18g (0.00227 mol) of solid PyBOP ([benzotriazolyloxy]tripyrrolidinophosphonium hexafluorophosphate, Advanced ChemTech) directly. Thereaction mixture was stirred for 30 minutes after which time it wasconcentrated in vacuo. The brown oil was treated with 10% aqueoushydrochloric acid. The suspension was extracted with ether. The organicextraction was washed with 10% sodium hydroxide followed by another 10%hydrochloric acid wash, was dried (MgSO₄) and concentrated in vacuo toafford 1.0 g of a light-brown foam. This intermediate was dissolved in25 mL of ethanolic hydrogen chloride, and the solution was allowed tostand at room temperature for 15 minutes. The reaction mixture wasconcentrated in vacuo to a brown oil that was purified by flash silicachromatography. Elution with dichloromethane→dichloromethane-methanol(166:1) afforded 0.2284 g of a light-brown viscous oil. Scratching withpentane-hexanes and drying under high vacuum afforded 0.1541 g (23%) ofan off-white foam; mp 61-75° C.;

¹H NMR (400 MHz, DMSO): δ 11.34 (s, 1H), 9.68 (s, 1H), 9.18 (s, 1H),7.65 (d, 1H, J=1.5 Hz), 7.58 (dd, 1H, J=8.7, 6.8 Hz), 7.52 (dd, 1H,J=8.4, 1.9 Hz), 7.15 (d, 1H, J=8.4 Hz), 6.74 (dd, 1H, J=11.8, 2.4 Hz),6.62 (ddd, 1H, J=8.4, 8.4, 2.7 Hz), 2.18 (s, 3H);

¹³C NMR (100 MHz, DMSO): δ 165.91, 164.36 (d, J_(C-F)=247.1 Hz), 146.78,139.18, 138.77, 135.43, 132.64, 130.60 (d, J_(C-F)=11.5 Hz), 122.23,112.52, 104.72 (d, J=22.1 Hz), 100.45 (d, J_(C-F)=25.2 Hz), 86.77,17.03;

¹⁹F NMR (376 MHz, DMSO): δ−107.20 to −107.27 (m);

IR (KBr) 3307 (broad, O-H stretch), 1636 (C═O stretch) cm⁻¹;

MS (CI) M+1=387.

Analysis calculated for C₁₄H₁₂FIN₂O₂:

C, 43.54; H, 3.13; N, 7.25.

Found: C, 43.62; H, 3.24; N, 6.98.

EXAMPLE 2

5-Bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

(a) Preparation of 5-Bromo-2,3,4-trifluorobenzoic acid

To a stirred solution comprised of 1-bromo-2,3,4-trifluorobenzene(Aldrich, 99%; 5.30 g, 0.0249 mol) in 95 mL of anhydrous tetrahydrofurancooled to −78° C. was slowly added 12.5 mL of 2.0 M lithiumdiisopropylamide in heptane/tetrahydrofuran/ethylbenzene solution(Aldrich). The mixture was stirred for 1 hour and transferred by canulainto 700 mL of a stirred saturated ethereal carbon dioxide solutioncooled to −78° C. The cold bath was removed, and the reaction mixturewas stirred for 18 hours at ambient temperature. Dilute (10%) aqueoushydrochloric acid (ca. 500 mL) was poured into the reaction mixture, andthe mixture was subsequently concentrated on a rotary evaporator to acrude solid. The solid product was partitioned between diethyl ether(150 mL) and aq. HCl (330 mL, pH 0). The aqueous phase was extractedwith a second portion (100 mL) of diethyl ether, and the combinedethereal extracts were washed with 5% aqueous sodium hydroxide (200 mL)and water (100 mL, pH 12). These combined alkaline aqueous extractionswere acidified to pH 0 with concentrated aqueous hydrochloric acid. Theresulting, suspension was extracted with ether (2×200 mL). The combinedorganic extracts were dried (MgSO₄), concentrated in vacuo, andsubjected to high vacuum until constant mass was achieved to afford 5.60g (88% yield) of an off-white powder; mp 139-142.5° C.;

¹H NMR (400 MHz, DMSO): δ 13.97 (broad s, 1H, 8.00-7.96 (m, 1H); ¹³C NMR(100 MHz, DMSO): δ 162.96, 129.34, 118.47, 104.54 (d, J_(C-F)=22.9 Hz);

¹⁹F NMR (376 MHz, DMSO): δ−120.20 to −120.31 (m), −131.75 to −131.86(m), −154.95 to −155.07 (m);

IR (KBr) 1696 (C═O stretch)cm⁻¹;

MS (CI) M+1=255.

Analysis calculated for C₇₄H₂₁BrF₃O₂:

C, 32.97; H, 0.79; N, 0.00; Br, 31.34; F, 22.35.

Found: C, 33.18; H, 0.64; N, 0.01; Br, 30.14; F, 22.75.

(b) Preparation of5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

To a stirred solution comprised of 1.88 g (0.00791 mol) of2-amino-5-iodotoluene in 10 mL of tetrahydrofuran at −78° C. was added 6mL (0.012 mol) of a 2.0 M lithium diisopropylamide intetrahydrofuran/heptane/ethylbenzene (Aldrich) solution. The resultinggreen suspension was stirred vigorously for 10 minutes, after which timea solution of 1.00 g (0.00392 mol) of 5-bromo-2,3,4-trifluorobenzoicacid in 15 mL of tetrahydrofuran was added. The cold bath wassubsequently removed, and the reaction mixture stirred for 18 hours. Themixture was concentrated, and the concentrate was treated with 100 mL ofdilute (10%) aqueous hydrochloric acid. The resulting suspension wasextracted with ether (2×150 mL), and the combined organic extractionswere dried (MgSO₄) and concentrated in vacuo to give an orange solid.The solid was triturated with boiling dichloromethane, cooled to ambienttemperature, and collected by filtration. The solid was rinsed withdichloromethane, and dried in the vacuum-oven (80° C.) to afford 1.39 g(76%) of a yellow-green powder; mp 259.5-262° C.;

¹H NMR (400 MHz, DMSO): δ 9.03 (s, 1H), 7.99 (dd, 1H, J=7.5, 1.9 Hz),7.57 (dd, 1H, J=1.5 Hz), 7.42 (dd, 1H, J=8.4, 1.9 Hz), 6.70 (dd, 1H,J=8.4, 6.0 Hz), 2.24 (s, 3H);

¹⁹F NMR (376 MHz, DMSO): δ−123.40 to −123.47 (m); −139.00 to −139.14(m);

IR (KBr) 1667 (C═O stretch)cm⁻¹;

MS (CI) M+1=469.

Analysis calculated for C₁₄H₉BrF₂INO₂:

C, 35.93; H, 1.94; N, 2.99; Br, 17.07; F, 8.12; I, 27.11.

Found: C, 36.15; H, 1.91; N, 2.70; Br, 16.40; F, 8.46; I, 26.05.

(c) Preparation of5-Bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

To a stirred solution comprised of5-bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid (0.51g, 0.0011 mol), O-(tetrahydro-2H-pyran-2-yl)-hydroxylamine (0.15 g,0.0013 mol), and diisopropylethylamine (0.25 mL, 0.0014 mol) in 20 mL ofan equivolume tetrahydrofuran-dichloromethane solution was added 0.6794g (0.001306 mol) of solid PyBOP (Advanced ChemTech) directly. Thereaction mixture was stirred at 24° C. for 10 minutes, and then wasconcentrated to dryness in vacuo. The concentrate was suspended in 100mL of 10% aqueous hydrochloric acid. The suspension was extracted with125 mL of diethyl ether. The ether layer was separated, washed with 75mL of 10% aqueous sodium hydroxide, and then with 100 mL of dilute acid.The ether solution was dried (MgSO₄) and concentrated in vacuo to afford0.62 g (100%) of an off-white foam. The foam was dissolved in ca. 15 mLof methanolic hydrogen chloride. After 5 minutes, the solution wasconcentrated in vacuo to an oil, and the oil was purified by flashsilica chromatography. Elution withdichloromethane→dichloromethane-methanol (99:1) afforded 0.2233 g (42%)of a yellow powder. The powder was dissolved in diethyl ether and washedwith dilute hydrochloric acid. The organic phase was dried (MgSO₄) andconcentrated in vacuo to afford 0.200 g of a foam. This product wastriturated with pentane to afford 0.1525 g of a powder that wasrepurified by flash silica chromatography. Elution with dichloromethaneafforded 0.0783 g (15%) of an analytically pure title compound, mp80-90° C.;

¹H NMR (400 MHz, DMSO): δ 11.53 (s, 1H), 9.38 (s, 1H), 8.82 (s, 1H),7.70 (dd, 1H, J=7.0, 1.9 Hz), 7.53 (s, 1H), 7.37 (dd, 1H, J=8.4, 1.9Hz), 6.55 (dd, 1H, J=8.2, 6.5 Hz), 2.22 (s, 3H);

¹⁹F NMR (376 MHz, DMSO): δ−126.24 to −126.29 (m), −137.71 to −137.77(m);

IR (KBr) 3346 (broad, O—H stretch), 1651 (C═O stretch)cm⁻¹;

MS (CI) M+1=484.

Analysis calculated for C₁₄H₁₀BrF₂IN₂O₂:

C, 34.81; H, 2.09; N, 5.80.

Found: C, 34.53; H, 1.73; N, 5.52,

Examples 3 to 12 and 78 to 102 in the table below were prepared by thegeneral procedures of Examples 1 and 2.

EXAMPLES 13-77

Examples 13 to 77 were prepared utilizing combinatorial syntheticmethodology by reacting appropriately substituted phenylamino benzoicacids (e.g., as shown in Scheme 1) and hydroxylamines

A general method is given below:

To a 0.8 mL autosampler vial in a metal block was added 40 μL of a 0.5 Msolution of the acid in DMF and 40 μL of the hydroxylamine (2 M solutionin Hunig's base and 1 M in amine in DMF). A 0.5 M solution of PyBrop wasfreshly prepared, and 50 μL were added to the autosampler vial. Thereaction was allowed to stand for 24 hours.

The reaction mixture was transferred to a 2 dram vial and diluted with 2mL of ethyl acetate. The organic layer was washed with 3 mL of distilledwater and the water layer washed again with 2 mL of ethyl acetate. Thecombined organic layers were allowed to evaporate to dryness in an openfume hood.

The residue was taken up in 2 mL of 50% acetonitrile in water andinjected on a semi-prep reversed phase column (10 mm×25 cm, 5 μMspherical silica, pore Size 115 A derivatised with C-18, the sample waseluted at 4.7 mL/min with a linear ramp to 100% acetonitrile over 8.5minutes. Elution with 100% acetonitrile continued for 8 minutes.)Fractions were collected by monitoring at 214 nM. The desired fractionswere evaporated using a Zymark Turbovap. The product was dissolved inchloroform and transferred to a preweighed vial, evaporated, and weighedagain to determine the yield. The structure was ed by mass spectroscopy.

EXAMPLES 3-102

Example Melting MS No. Compound Point (° C.) (M-H⁺) 32-(4-bromo-2-methyl-phenylamino)-4-fluoro-N-    56-75 dec 523hydroxy-benzamide 4 5-Chloro-N-hydroxy-2-(4-iodo-2-methyl- 65 decphenylamino)-benzamide 5 5-Chloro-N-hydroxy-2-(4-iodo-2-methyl- 62-67phenylamino)-N-methyl-benzamide 65-Chloro-2-(4-iodo-2-methyl-phenylamino)-N- 105-108(terahydropyran-2-yloxy)benzamide 75-Chloro-2-(4-iodo-2-methyl-phenylamino)-N- 64-68 methoxybenzamide 84-Fluoro-N-hydroxy-2-(4-fluoro-2-methyl- 119-135 phenylamino)-benzamide9 4-Fluoro-N-hydroxy-2-(2-methyl phenylamino)- 101-103 benzamide 104-Fluoro-2-(4-fluor-2-methyl-phenylamino)-N- 142-146(terahydropyran-2-yloxy)benzamide 114-Fluoro-N-hydroxy-2-(4-cluoro-2-methyl- 133.5-135  phenylamino)-benzamide 12 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-  107-109.5 phenylmethoxy-benzamide 134-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 399 methoxy-benzamide 143,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 417 N-methoxy-benzamide 152-(4-Bromo-2-methyl-phenylamino)- 369 3,4-difluoro-N-methoxy-benzamide16 2-(4-Bromo-2-methyl-phenylamino)-N-ethoxy-  342*3,4-difluoro-benzamide (M-EtO) 175-Bromo-N-ethoxy-3,4-difluoro-2-(4-iodo- 5092-methyl-phenylamino)-benzamide 183,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 445 N-isopropoxy-benzamide19 2-(4-Bromo-2-methyl-phenylamino)- 3973,4-difluoro-N-isopropoxy-benzamide 204-Fluoro-N-(furan-3-ylmethoxy)-2-(4-iodo- 4652-methyl-phenylamino)-benzamide 213,4-Difluoro-N-(furan-3-ylmethoxy)-2-(4-iodo- 4832-methyl-phenylamino)-benzamide 22 2-(4-Bromo-2-methyl-phenylamino) 4353,4-difluoro-N-(furan-3-ylmethoxy)-benzamide 235-Bromo-3,4-difluoro-N-(furan-3-ylmethoxy)- 5612-(4-iodo-2-methyl-phenylamino)-benzamide 245-Bromo-N-(but-2-enyloxy)-3,4-difluoro- 5362-(4-iodo-2-methyl-phenylamino)-benzamide 254-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 423(prop-2-ynyloxy)-benzamide 263,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 441N-(prop-2-ynyloxy)-benzamide 273,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 455N-(1-methyl-prop-2-ynyloxy)-benzamide 282-(4-Bromo-2-methyl-phenylamino)- 4073,4-difluoro-N-(1-methyl-prop-2-ynyloxy)- benzamide 29N-(But-3-ynyloxy)-3,4-difluoro-2-(4-iodo- 4552-methyl-phenylamino)-benzamide 302-(4-Bromo-2-methyl-phenylamino)-N-(but- 4073-ynyloxy)-3,4-difluoro-benzamide 315-Bromo-N-(but-3-ynyloxy)-3,4-difluoro- 5332-(4-iodo-2-methyl-phenylamino)-benzamide 323,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 517N-(3-phenyl-prop-2-ynyloxy)-benzamide 333,4-Difluoro-2-(4-bromo-2-methyl- 469phenylamino)-N-(3-phenyl-prop-2-ynyloxy)- benzamide 343,4-Difluoro-N-[3-(3-fluoro-phenyl)-prop- 5352-ynyloxy]-2-(4-iodo-2-methyl-phenylamino)- benzamide 352-(4-Bromo-2-methyl-phenylamino)- 4873,4-difluoro-N-[3-(3-fluoro-phenyl)-prop- 2-ynyloxy]-benzamide 363,4-Difluoro-N-[3-(2-fluoro-phenyl)-prop- 5352-ynyloxy]-2-(4-iodo-2-methyl-phenylamino)- benzamide 375-Bromo-3,4-difluoro-N-[3-(2-fluoro-phenyl)- 613prop-2-ynyloxy]-2-(4-iodo-2-methyl- phenylamino)-benzamide 392-(4-Bromo-2-methyl-phenylamino)- 5103,4-difluoro-N-(3-methyl-5-phenyl-pent-2-en- 4-ynyloxy)-benzamide 40N-Ethoxy-3,4-difluoro-2-(4-iodo-2-methyl- 431 phenylamino)-benzamide 412-(4-Bromo-2-methyl-phenylamino)-N-ethoxy- 383 3,4-difluoro-benzamide 424-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 427 propoxy-benzamide 433,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 445 N-propoxy-benzamide 442-(4-Bromo-2-methyl-phenylamino)- 397 3,4-difluoro-N-propoxy-benzamide45 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 523phenylamino)-N-propoxy-benzamide 464-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 427 isopropoxy-benzamide 473,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 445 N-isopropoxy-benzamide48 2-(4-Bromo-2-methyl-phenylamino)- 3973,4-difluoro-N-isopropoxy-benzamide 495-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 523phenylamino)-N-isopropoxy-benzamide 50N-Cyclobutyloxy-3,4-difluoro-2-(4-iodo- 4572-methyl-phenylamino)-benzamide 51 2-(4-Bromo-2-methyl-phenylamino)-N-409 cyclobutyloxy-3,4-difluoro-benzamide 52N-Cyclopentyloxy-4-fluoro-2-(4-iodo-2-methyl- 453 phenylamino)-benzamide53 N-Cyclopentyloxy-3,4-difluoro-2-(4-iodo- 4712-methyl-phenylamino)-benzamide 54 2-(4-Bromo-2-methyl-phenylamino)-N-423 cyclopentyloxy-3,4-difluoro-benzamide 55N-Cyclopropylmethoxy-4-fluoro-2-(4-iodo- 4392-methyl-phenylamino)-benzamide 56N-Cyclopropylmethoxy-3,4-difluoro-2-(4-iodo- 4572-methyl-phenylamino)-benzamide 57 2-(4-Bromo-2-methyl-phenylamino)-N-409 cyclopropylmethoxy-3,4-difluoro-benzamide 585-Bromo-N-cyclopropylmethoxy-3,4-difluoro- 4352-(4-iodo-2-methyl-phenylamino) 594-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 505(2-phenoxy-ethoxy)-benzamide 603,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 523N-(2-phenoxy-ethoxy)-benzamide 61 2-(4-Bromo-2-methyl-phenylamino)- 4753,4-difluoro-N-(2-phenoxy-ethoxy)-benzamide 624-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 481(thiophen-2-ylmethoxy)-benzamide 633,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 499N-(thiophen-2-ylmethoxy)-benzamide 64 2-(4-Bromo-2-methyl-phenylamino)-451 3,4-difluoro-N-(thiophen-2-ylmethoxy)- benzamide 654-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 439(2-methyl-allyloxy)-benzamide 663,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 457N-(2-methyl-allyloxy)-benzamide 67 2-(4-Bromo-2-methyl-phenylamino)- 4103,4-difluoro-N-(2-methyl-allyloxy)-benzamide 68N-(But-2-enyloxy)-4-fluoro-2-(4-iodo-2-methyl- 439phenylamino)-benzamide 69 N-(But-2-enyloxy)-3,4-difluoro-2-(4-iodo- 4572-methyl-phenylamino)-benzamide 702-(4-Bromo-2-methyl-phenylamino)-N-(but- 4102-enyloxy)-3,4-difluoro-benzamide 713,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 441N-(prop-2-ynyloxy)-benzamide 72N-(But-3-ynyloxy)-3,4-difluoro-2-(4-iodo- 4552-methyl-phenylamino)-benzamide 73 2-(4-Bromo-2-methyl-phenylamino)-N-449 (4,4-dimethyl-pent-2-ynyloxy)-3,4-difluoro- benzamide 74N-(But-2-enyloxy)-3,4-difluoro-2-(4-iodo- 4572-methyl-phenylamino)-benzamide 752-(4-Bromo-2-methyl-phenylamino)-N-(but- 4102-enyloxy)-3,4-difluoro-benzamide 76N-(3-tert.-butyl-propyn-2-yl)oxy-4-fluoro- 4792-(4-iodo-2-methyl-phenylamino)-benzamide 774-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-  577*phenylmethoxy-benzamide *CI 78 4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-oil phenylamino)-N-isopropyl-benzamide 79N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(4- 125-127iodo-2-methyl-phenylamino)-benzamide 804-Fluoro-N-hydroxy-2-(4-iodo-2-methyl- 45-55phenylamino)-N-methyl-benzamide 814-Fluoro-N-hydroxy-2-(4-iodo-2-methyl- 208-209phenylamino)-5-nitro-benzamide (GLASS) 822-(2-Chloro-4-iodo-phenylamino)-N-hydroxy-4- 199-200 nitro-benzamide 833,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 163-165N-(tetrahydro-pyran-2-yloxy)-benzamide 843,4-Difluoro-N-hydroxy-2-(4-iodo-2-methyl- 65-75 phenylamino)-benzamide85 3,4-Difluoro-5-bromo-2-(4-iodo-2-methyl- 95phenylamino)-N-(2-piperidin-1-yl-ethoxy)- benzamide 865-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 167-169phenylamino)-N-(tetrahydro-pyran-2-yloxy)- benzamide 872-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N- 165-169 hydroxy-benzamide(HCl salt) 88 2-(2-Chloro-4-iodo-phenylamino)-4-fluoro-N-   166-167.5(tetrahydro-pyran-2-yloxy)-benzamide 893,4-Difluoro-2-(2-chloro-4-iodo-phenylamino)- 173-174N-cyclobutylmethoxy-benzamide 903,4-Difluoro-2-(2-chloro-4-iodo-phenylamino)- 121-122N-(tetrahydro-pyran-2-yloxy)-benzamide 915-Bromo-2-(2-chloro-4-iodo-phenylamino)-N-   206-211.5(2-dimethylamino-ethoxy)-3,4-difluoro- DEC benzamide monohydrochloridesalt 92 5-Bromo-N-(2-dimethylamino-propoxy)-3,4-  95-105difluoro-2-(4-iodo-2-methyl-phenylamino)- benzamide 935-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4- 266-280difluoro-N-hydroxy-benzamide DEC 945-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4- 167.5-169.5difluoro-N-(tetrahydro-pyran-2-yloxy)- benzamide 953,4-Difluoro-2-(2-chloro-4-iodo-phenylamino)- 172.5-173.5N-cyclopropylmethoxy-benzamide 965-Bromo-2-(2-chloro-4-iodo-phenylamino)-N-   171-172.5cyclopropylmethoxy-3,4-difluoro-benzamide 975-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 173.5-175  phenylamino)-N-(2-morpholin-4-yl-ethoxy)- benzamide 985-Bromo-N-(2-diethylamino-ethoxy)-3,4- 81 DECdifluoro-(4-iodo-2-methyl-phenylamino)- benzamide 995-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 126-128phenylamino)-N-isobutoxy-benzamide 1005-Bromo-N-cyclohexylmethoxy-3,4-difluoro-2- 139-142(4-iodo-2-methyl-phenylamino)-benzamide 1015-Bromo-N-cyclopentylmethoxy-3,4-difluoro-2- 113-115(4-iodo-2-methyl-phenylamino)-benzamide 1025-Bromo-N-cyclobutylmethoxy-3,4-difluoro-2- 138-139(4-iodo-2-methyl-phenylamino)-benzamide

The invention compounds are useful in treating cancer and otherproliferative diseases by virtue of their selective inhibition of thedual specificity protein kinases MEK₁ and MEK₂. The invention compoundhas been evaluated in a number of biological assays which are normallyutilized to establish inhibition of proteins and kinases, and to measuremitogenic and metabolic responses to such inhibition.

Enzyme Assays

Cascade Assay for Inhibitors of the MAP Kinase Pathway

Incorporation of ³²P into myelin basic protein (MBP) was assayed in thepresence of a glutathione S-transferase fusion protein containing p44MAPkinase (GST-MAPK) and a glutathione S-transferase fusion proteincontaining p45MEK (GST-MEK). The assay solution contained 20 mM HEPES,pH 7.4, 10 mM MgCl₂, 1 mM MnCl₂, 1 mM EGTA, 50 μM [γ-³²P]ATP, 10 μgGST-MEK, 0.5 μg GST-MAPK and 40 μg MBP in a final volume of 100 μL.Reactions were stopped after 20 minutes by addition of trichloroaceticacid and filtered through a GF/C filter mat. ³²P retained on the filtermat was determined using a 1205 Betaplate. Compounds were assessed at 10μM for ability to inhibit incorporation of ³²P.

To ascertain whether compounds were inhibiting GST-MEK or GST MAPK, twoadditional protocols were employed. In the first protocol, compoundswere added to tubes containing GST-MEK, followed by addition ofGST-MAPK, MBP and [γ-³²P]ATP. In the second protocol, compounds wereadded to tubes containing both GST-MEK and GST-MAPK, followed by MBP and[γ-³²P]ATP. Compounds that showed activity in both protocols were scoredas MAPK inhibitors, while compounds showing activity in only the firstprotocol were scored as MEK inhibitors.

In Vitro MAP Kinase Assay

Inhibitory activity was also confirmed in direct assays. For MAP kinase,1 μg GST-MAPK was incubated with 40 μg MBP for 15 minutes at 30° C. in afinal volume of 50 μL containing 50 mM Tris (pH 7.5), 10 μM MgCl₂, 2 μMEGTA, and 10 μM [γ-³²P]ATP. The reaction was stopped by addition ofLaemmli SDS sample buffer and phosphorylated MBP resolved byelectrophoresis on a 10% polyacrylamide gel. Radioactivity incorporatedinto MBP was determined by autoradiography, and subsequently by excisionof the bands followed by scintillation counting.

In Vitro MEK Assay

For evaluation of direct MEK activity, 10 μg GST-MEK1 was incubated with5 μg of a glutathione S-transferase fusion protein containing p44MAPkinase with a lysine to alanine mutation at position 71 (GST-MAPK-KA).This mutation eliminates kinase activity of MAPK, so only kinaseactivity attributed to the added MEK remains. Incubations were 15minutes at 30° C. in a final volume of 50 μL containing 50 mM Tris (pH7.5), 10 μM MgCl₂, 2 μM EGTA, and 10 μM [γ-³²P]ATP. The reaction wasstopped by addition of Laemmli SDS sample buffer and phosphorylatedGST-MAPK-KA was resolved by electrophoresis on a 10% polyacrylamide gel.Radioactivity incorporated into GST-MAPK-KA was determined byautoradiography, and subsequently by excision of the bands followed byscintillation counting. Additionally, an artificially activated MEK wasutilized that contained serine to glutamate mutations at positions 218and 222 (GST-MEK-2E). When these sites are phosphorylated, MEK activityis increased. Phosphorylation of these sites can be mimicked by mutationof the serine residues to glutamate. For this assay, 5 μg GST-MEK-2E wasincubated with 5 μg GST-MAPK-KA for 15 minutes at 30° C. in the samereaction buffer as described above. Reactions were terminated andanalyzed as above.

Whole Cell MAP Kinase Assay

To determine if compounds were able to block activation of MAP kinase inwhole cells, the following protocol was used: Cells were plated inmulti-well plates and grown to confluence. Cells were thenserum-deprived overnight. Cells were exposed to the desiredconcentrations of compound or vehicle (DMSO) for 30 minutes, followed byaddition of a growth factor, eg, PDGF (100 ng/mL). After a 5-minutetreatment with the growth factor, cells were washed with PBS, then lysedin a buffer consisting of 70 mM NaCl, 10 mM HEPES (pH 7.4), 50 mMglycerol phosphate, and 1% Triton X-100. Lysates were clarified bycentrifugation at 13,000×g for 10 minutes. Five micrograms of theresulting supernatants were incubated with 10 μg microtubule associatedprotein-2 (Map2) for 15 minutes at 30° C. in a final volume of 25 μLcontaining 50 mM Tris (pH 7.4), 10 mM MgCl₂, 2 mM EGTA and 30 μM[γ-³²P]ATP. Reactions were terminated by addition of Laemmli samplebuffer. Phosphorylated Map2 was resolved on 7.5% acrylamide gels andincorporated radioactivity determined by autoradiography and subsequentexcision of the bands followed by scintillation counting.

Immunoprecipitation and Antiphosphotyrosine Immunoblots

To determine the state of tyrosine phosphorylation of cellular MAPkinase, cells were lysed, endogenous MAP kinase was immunoprecipitatedwith a specific antibody, and the resulting immunoprecipitate analyzedfor the presence of phosphotyrosine as follows: confluent cells wereserum-deprived overnight and treated with compounds and growth factorsas described above. Cells were then scraped and pelleted at 13,000×g for2 minutes. The resulting cell pellet was resuspended and dissolved in100 μL of 1% SDS containing 1 mM NaVO₄. Following alternate boiling andvortexing to denature cellular protein, 900 μL RIPA buffer (50 mM Tris(pH 7.4), 150 mM NaCl, 1% Triton X-100, 0.1% deoxycholate, and 10 mMEDTA) was added. To this mixture was added 60 μL agarose beads coupledwith rabbit immunoglobulin G and 60 μL Pansorbin cells in order to clearthe lysate of nonspecific binding proteins. This mixture was incubatedat 4° C. for 15 minutes then centrifuged at 13,000×g for 10 minutes. Theresulting supernatant was transferred to fresh tubes and incubated with10 μL of a polyclonal antisera raised against a fragment of MAP kinasefor a minimum of 1 hour at 4° C. Seventy microliters of a slurry ofagarose beads coupled with protein G and protein A was added and theincubation continued for an additional 30 minutes at 4° C. The beadswere pelleted by centrifugation at 13,000×g for 5 minutes and washedthree times with 1 mL RIPA buffer. Laemmli sample buffer was added tothe final bead pellet. This mixture was boiled for 5 minutes thenresolved on a 10% acrylamide gel. Proteins on the gel were transferredto a nitrocellulose membrane and nonspecific binding sites on themembrane blocked by incubation with 1% ovalbumin and 1% bovine serumalbumin in TBST (150 mM NaCl, 10 mM Tris (pH 7.4), and 0.05% Tween 20).The membrane was then incubated with a commercially available antibodydirected against phosphotyrosine. Antibody bound on the membrane wasdetected by incubation with ¹²⁵I-protein A, followed by autoradiography.

Cell Growth Assays

³H-Thymidine Incorporation

Cells were plated in multi-well plates and grown to near confluence. Themedia was then removed and replaced with growth media containing 1%bovine serum albumin. After 24-hour serum starvation, compounds andspecific growth factors were added and incubations continued for anadditional 24 hours. During the final 2 hours, ³H-thymidine was added tothe medium. To terminate the incubations, the medium was removed andcell layers washed twice with ice-cold phosphate-buffered saline. Afterthe final wash, ice-cold 5% trichloroacetic acid was added and the cellsincubated for 15 minutes at room temperature. The trichloroacetic acidsolution was then removed and the cell layer washed three times withdistilled water. After the final wash, the cell layer was solubilized byaddition of 2% sodium dodecylsulfate. Radioactivity in this solution wasdetermined by scintillation counting.

In 3T3-L1 adipocyte cells, in which the inhibition blocks MAPKactivation by insulin with an IC₅₀ of 3 μM, the compound had no effecton the insulin stimulated uptake of radiolabeled 2-deoxyglucose, or onthe insulin-stimulated synthesis of either lipid or glycogen at 10 μMconcentration. This demonstrates that the inhibitor shows selectivitybetween the mitogenic and metabolic effects of insulin, and demonstratesthat the inhibitor will show less toxicity than an inhibitor which doesnot show this surprising selectivity.

Monolayer Growth

Cells were plated into multi-well plates at 10 to 20,000 cells/mL.Forty-eight hours after seeding, compounds were added to the cell growthmedium and incubation was continued for 2 additional days. Cells werethen removed from the wells by incubation with trypsin and enumeratedwith a Coulter counter.

Growth in Soft-agar

Cells were seeded into 35-mm dishes at 5 to 10,000 cells/dish usinggrowth medium containing 0.3% agar. After chilling to solidify the agar,cells were transferred to a 37° C. incubator. After 7 to 10 days growth,visible colonies were manually enumerated with the aid of a dissectingmicroscope.

Order of addition experiments established that the invention compoundsare inhibiting MEK and not MAP kinase. Experiments looking at thephosphorylation of a kinase defective mutant of MAP kinase as substrate(so that there can be no autophosphorylation of the MAP kinase tocomplicate interpretation) confirms that the inhibitor inhibits MEK withan IC₅₀ essentially identical to that produced in the cascade assay.

Kinetic analysis demonstrates that the invention compounds are notcompetitive with ATP. Thus, they do not bind at the ATP binding site ofthe enzyme, which is probably the explanation as to why these compoundsdo not show the nonspecific kinase inhibitory activity typical of mostkinase inhibitors, which do bind at the ATP binding site and which areATP competitive.

The in vitro and in vivo biological activity of several representativecompounds of Formula I in the foregoing assays is presented in Table 1.Data for several known compounds is also presented.

TABLE I In vivo Compound of In vitro (cell culture) Example No. IC₅₀(μM) IC₅₀ (μM) 1 0.007 0.05 2 0.003 0.03 3 0.072 3 4 0.023 1 5 0.566 ˜306 0.345 ˜30 7 0.221 <30 8 7.13 3 9 0.409 1 11 0.334 0.5 12 0.826 130.243 14 0.061 >2 17 0.014 20 0.042 0.17 21 0.014 22 0.137 23 0.016 240.021 0.12 25 0.102 27 0.026 28 0.728 29 0.076 0.73 30 0.971 31 0.045 320.017 33 0.374 34 0.113 1.5 36 0.056 0.07 37 0.002 38 0.077 0.065 390.147 40 0.028 0.125 41 0.236 42 0.087 43 0.040 0.100 44 0.475 45 0.12647 0.087 0.13 49 0.085 50 0.043 0.22 53 0.140 55 0.047 56 0.014 57 0.18158 0.018 0.014 59 0.259 62 0.086 63 0.019 64 0.279 65 0.057 66 0.0160.13 68 0.119 69 0.016 70 0.224 71 0.015 0.39 74 0.035 77 0.28 78 0.08079 0.008 80 0.080 81 0.017 82 0.003 0.04 83 0.031 84 0.001 0.005 850.024 86 0.047 87 <0.001 88 0.069 89 0.005 0.30 90 0.055 91 0.020 920.033 93 0.010 0.05 94 0.038 95 0.001 96 <0.010 97 0.015 98 0.025 990.018 0.50 100 0.026 >1 101 0.008 >1 102 0.004 0.20

The following compounds, which are disclosed in U.S. Pat. No. 5,155,110,were also evaluated in the foregoing assays, and each such compounddemonstrated little or no inhibitory activity.

% Inhibition R₆ R₇ In Vitro 

H H 9 at 1 μM −3 at 10 μM H CH₃ −8 at 1 μM 8 at 10 μM CH₃ H −5 at 1 μM19 at 10 μM iPr H 17 at 1 μM 9 at 10 μM CH₂Ph H −4 at 1 μM 18 at 10 μM

H H 6 at 1 μM −4 at 10 μM H CH₃ −6 at 1 μM 12 at 10 μM CH₃ H 13 at 1 μM19 at 10 μM iPr H −11 at 1 μM 7 at 10 μM

EXAMPLE 103

The compound from Example 95,2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluorobenzamide,was evaluated in animals implanted with a murine colon tumor, C26/clone10. Male CD2F1 mice (NCI: Charles River, Kingston) were implantedsubcutaneously with tumor fragments (approximately 30 mg) in the regionof the right axilia on Day 0. The compound of Example 95 wasadministered intraperitoneally (IP) or orally (PO) on Days 1 through 14,post-implant, for a total of 14 days (6 mice per group). The vehicle forthe test compound, and for control animals, was 10% EtOH/10%Cremophor-EL (Sigma)/80% H₂O, pH 5.0. Tumor volumes were recorded threetimes per week by measuring the length and width of the individualtumors and calculating mass in milligrams according to the formula(a×b²)/2, where a and b are the length and width of the tumor. Percenttreated/control (T/C) was calculated based on the ratio of the mediantumor volume of the treated tumors compared with the median tumor volumeof control animals on specified measurement days.

In the trial in which the compound of Example 95 was administered IP,the doses were 200, 124, 77, and 48 mg/kg/day. The invention compoundinhibited tumor growth by 59% to 100% as assessed on Day 15. The mediansize of the control tumors on Day 15 was 1594 mg. Table 2 shows thenumber of animal deaths in each treatment group, the change in bodyweight, the percent of the median tumor volume of the treated groupcompared to the control group, and the percent inhibition.

TABLE 2 Change in Body Weight % T/C Dose Non-Specific Deaths (grams)(Day 15) % Inhibition 200 1/6 +2 0 100 124 1/6 +3 4 96 77 2/5 +2 2 98 480/6 +3 41 59

In the test in which the compound of Example 95 was orally administered,the doses were 300, 186, 115, and 71 mg/kg/day. The invention compoundinhibited tumor growth 64% to 83% as assessed on Day 17. The median sizeof the control tumors on Day 17 was 1664 mg. Table 3 shows the number ofanimal deaths in each treatment group, the change in body weight, thepercent of the median tumor volume of the treated group compared to thecontrol group, and the percent inhibition.

TABLE 3 Change in Body Weight % T/C Dose Non-Specific Deaths (grams)(Day 15) % Inhibition 300 0/6 +2 17 83 186 0/6 +2 25 75 115 1/6 +2 21 7971 0/6 +2 36 64

The foregoing assay established that the invention compounds of FormulaI are particularly useful for treating cancers such as colon cancer. Thecompounds are especially well-suited for use in combination withradiation to treat and control cancers.

The invention compounds will be utilized to treat subjects sufferingfrom cancer and other proliferative diseases and in need of treatment.The compounds are ideally suited to treating psoriasis, restenosis,autoimmune disease, and atherosclerosis. The compounds will generally beutilized as a pharmaceutical formulation, in which the compound ofFormula I is present in a concentration of about 5% to about 95% byweight. The compounds can be formulated for convenient oral, parenteral,topical, rectal, or like routes of administration. The compound will beformulated with common diluents, excipients, and carriers routinelyutilized in medicine, for instance, with polyols such as glycerin,ethylene glycol, sorbitol 70; mono- and difatty acid esters of ethyleneglycol. Starches and sugars such as corn starch, sucrose, lactose, andthe like, can be utilized for solid preparations. Such solidformulations can be in the form of tablets, troches, pills, capsules,and the like. Flavoring agents such as peppermint, oil of wintergreen,and the like can be incorporated.

Typical doses of active compound are those that are effective to treatthe cancer or other proliferative disorder afflicting the mammal. Doseswill generally be from about 0.1 mg per kilogram body weight to about500 mg per kilogram body weight. Such doses will be administered fromone to about four times a day, or as needed to effectively treat thecancer, psoriasis, restenosis, or other proliferative disorder.

A preferred method for delivering the invention compound is orally via atablet, capsule, solution, or syrup. Another method is parenterally,especially via intravenous infusion of a solution of the benzopyran inisotonic saline or 5% aqueous glucose.

Following are typical formulations provided by the invention.

EXAMPLE 104

Preparation of 50-mg Tablets Per 10,000 Per Tablet Tablets 0.050 g4-fluoro-N-hydroxy-2-(4-iodo-2-methyl- 500 g phenylamino)-benzamide0.080 g lactose 800 g 0.010 g corn starch (for mix) 100 g 0.008 g cornstarch (for paste) 80 g 0.002 g magnesium stearate (1%) 20 g 0.150 g1500 g

The benzhydroxamic acid, lactose, and corn starch (for mix) are blendedto uniformity. The corn starch (for paste) is suspended in 600 mL ofwater and heated with stirring to form a paste. The paste is used togranulate the mixed powders. The granules are passed through a #8 screenand dried at 120° F. The dry granules are passed through a #16 screen.The mixture is lubricated with 1% magnesium stearate and compressed intotablets. The tablets are administered to a mammal for inhibiting MEKenzymes and treating restenosis, atherosclerosis, and psoriasis.

EXAMPLE 105

Preparation of Oral Suspension Ingredient Amount5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N- 500 mg (methoxy)-benzamideSorbitol solution (70% NF) 40 mL Sodium benzoate 150 mg Saccharin 10 mgRed dye 10 mg Cherry flavor 50 mg Distilled water qs ad 100 mL

The sorbitol solution is added to 40 mL of distilled water and thebenzhydroxamic acid derivative is suspended therein. The saccharin,sodium benzoate, flavor, and dye are added and dissolved. The volume isadjusted to 100 mL with distilled water. Each milliliter of syrupcontains 5 mg of the invention compound. The syrup is administered to amammal for treating proliferative disease, especially breast cancer andskin cancer.

EXAMPLE 106

Preparation of Parenteral Solution

In a solution of 700 mL of propylene glycol and 200 mL of water forinjection is added 20.0 g of4-fluoro-2-(4-bromo-2-methyl-phenylamino)-N-(hydroxy)-benzamide. Thevolume of the solution is adjusted to 1000 mL by addition of water forinjection. The formulation is heat sterilized, filled into 50-mLampoules each containing 2.0 mL (40 mg of4-fluoro-2-(4-bromo-2-methyl-phenylamino)-N-(hydroxy)-benzamide), andsealed under nitrogen.

The invention compounds thus formulated will be administered to a mammalin need of treatment for a proliferative disorder such as cancer,psoriasis, restenosis, atherosclerosis, and autoimmune disease at a rateand dose effective to treat the condition. An “antiproliferative amount”of an invention compound is that quantity of compound that inhibits orreduces the rate of proliferation of target cells. Typical cancers to betreated according to this invention include breast cancer, colon cancer,prostate cancer, skin cancer, and the like. The compound is well-suitedto the treatment of psoriasis, restenosis, and atherosclerosis, and toinhibiting the activity of MEK enzymes, especially MEK₁ and MEK₂. Allthat is required is to administer to a mammal an MEK inhibiting amountof a compound of the invention. An “MEK inhibiting amount” of aninvention compound is an amount that when administered to a mammalcauses a measurable inhibition of the MEK enzyme. Typical MEK inhibitingamounts will be from about 0.1 μg to about 500 mg of active compound perkilogram body weight. For treating the proliferative diseases mentionedabove, typical doses will be from about 0.1 to about 50 mg/kg, normallygiven from one to about four times per day.

What is claimed is:
 1. A compound of Formula I

wherein: R₁ is hydrogen, hydroxy, C₁-C₈ alkyl, C₁-C₈ alkoxy, halo,trifluoromethyl, or CN; R₂ is hydrogen; R₃, R₄, and R₅ independently arehydrogen, hydroxy, halo, trifluoromethyl, C₁-C₈ alkyl, C₁-C₈ alkoxy,nitro, CN, or (O or NH)_(m)—(CH₂)_(n)—R₉, where R₉ is hydrogen, hydroxy,CO₂H or NR₁₀R₁₁; n is 0 to 4; m is 0 or 1; R₁₀ and R₁₁ independently arehydrogen or C₁-C₈ alkyl, or taken together with the nitrogen to whichthey are attached can complete a 3- to 10-member cyclic ring optionallycontaining one, two, or three additional heteroatoms independentlyselected from O, S, NH, and N—C₁-C₈ alkyl; R₆ is hydrogen, C₁-C₈ alkyl,

 aryl, aralkyl, or C₃-C₁₀ cycloalkyl; R₇ is cycloalkyl group having from2 to 9 carbon atoms and containing a NR₉ heteroatom; and wherein any ofthe foregoing alkyl, alkenyl, and alkynyl groups can be unsubstituted orsubstituted by cycloalkyl (or cycloalkyl optionally containing aheteroatom independently selected from O, S, and NR₉), aryl, aryloxy,heteroaryl, heteroaryloxy, or NR₁₀R₁₁.
 2. A compound according to claim1 wherein R₁ is C₁-C₈ alkyl or halo.
 3. A compound according to claim 2wherein R₆ is hydrogen.
 4. A compound according to claim 3 wherein R₁ ismethyl.
 5. A compound according to claim 4 having the formula


6. A compound of claim 5 wherein R₄ is fluoro, and R₃ and R₅ arehydrogen.
 7. A compound of claim 5 wherein R₃ and R₄ are fluoro, and R₅is hydrogen.
 8. A compound of claim 5 wherein R₃ and R₄ are fluoro, andR₅ is bromo.
 9. A compound according to claim 8 which is:5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-1-yl-ethoxy)-benzamide.10. A compound of claim 5 wherein R₃ and R₄ are hydrogen, and R₅ ishalo.
 11. A compound of claim 4 having the formula


12. A compound of claim 11 wherein R₃ and R₄ are fluoro, and R₅ ishydrogen.
 13. A pharmaceutical formulation comprising a compound ofclaim 1 admixed with a pharmaceutically acceptable excipient, diluent,or carrier.
 14. A formulation of claim 13 comprising a compound of theformula


15. A formulation of claim 13 comprising a compound of the formula


16. A method for treating a mammal suffering from psoriasis and in needof treatment comprising administering an effective amount of a compoundof claim
 1. 17. A method for treating a mammal suffering from breastcancer, colon cancer, prostate cancer, skin cancer or pancreatic cancerand in need of treatment, the method comprising administering aneffective amount of a compound of claim
 1. 18. A compound of claim 1having the formula


19. A compound of claim 1 having the formula


20. A compound of claim 1 having the formula


21. A compound of claim 1 having the formula